Nothing is currently known regarding gene regulation in bdellovibrios. A global analysis of gene regulation during the bdellovibrio developmental cycle will provide valuable information for later examination of individual genes. The overall objectives are to identify proteins being synthesized at various points of the life cycle of two distinct strains, to separate these proteins using gel electrophoresis, and to determine the kinetics of synthesis of selected proteins throughout the developmental cycle. Synchronously growing cultures of Bdellovibrio bacteriovorous 1O9JS or Bdellovibrio sp. strain Wr, will be pulse-labeled with 35s- methionine. Proteins will be acid precipitated and separated using high resolution two-dimensional gel electrophoresis. Labeled proteins will be visualized by autoradiography. Proteins which show increased or decreased levels of synthesis will be examined further. The labeled proteins will be cut out of gels and solubilized. Relative rates of synthesis of these proteins will be determined using the ratio of the amount of 35S in the spot to the total amount of 35S on the gel. By determining the rates of synthesis at many time points, the kinetic patterns can be determined. The cellular locations of these proteins will be determined by separating the inner and outer membranes from the soluble cytoplasmic fractions using detergents and differential centrifugation. Proteins in these fractions will then be examined by two-dimensional gel electrophoresis.